LOCUS       sequence-214250-        6585 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of His-Eos-I3 in insect cells (Baculovirus system).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6585)
  AUTHORS   Weith M, Seiler J, van den Boom J, Kracht M, Hulsmann J, Primorac I,
            Del Pino Garcia J, Kaschani F, Kaiser M, Musacchio A, Bollen M, 
            Meyer H
  TITLE     Ubiquitin-Independent Disassembly by a p97 AAA-ATPase Complex Drives
            PP1 Holoenzyme Formation.
  JOURNAL   Mol Cell. 2018 Oct 11. pii: S1097-2765(18)30791-3. doi: 
            10.1016/j.molcel.2018.09.020.
  PUBMED    30344098
REFERENCE   2  (bases 1 to 6585)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell. 
            2018 Oct 11. pii: S1097-2765(18)30791-3. doi: 
            10.1016/j.molcel.2018.09.020."
COMMENT     Sequence Label: pFL  His-mEos3.2-I3
FEATURES             Location/Qualifiers
     source          1..6585
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      202..656
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(289..308)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     498..519
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        682..786
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(787..855,856..1647)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1005..1024)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1818..2406
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2307..2326
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2560..2577
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2711..2935
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(3002..3535)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3724..3729,3730..3746,3747..3752))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(4148..4181)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    4371..4419
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4444..4463
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(4555..4664)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        4709..4800
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     4725..4742
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             4861..4878
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4900..4920
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             4933..5610
                     /codon_start=1
                     /product="improved monomeric variant of green-to-red 
                     photoswitchable fluorescent protein EosFP (Zhang et al., 
                     2012)"
                     /label=mEos3.2
                     /translation="MSAIKPDMKIKLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVK
                     EGGPLPFAFDILTTAFHYGNRVFAKYPDNIQDYFKQSFPKGYSWERSLTFEDGGICNAR
                     NDITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIEMALLL
                     EGNAHYRCDFRTTYKAKEKGVKLPGAHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
     polyA_signal    6194..6328
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6231..6250)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6285..6304
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  join(6438..6585,1..18)
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 ctattttgtc cgcccacagc ggggcatttt tcttcctgtt atgtttttaa tcaaacatcc
       61 tgccaactcc atgtgacaaa ccgtcatctt cggctacttt ttctctgtca cagaatgaaa
      121 atttttctgt catctcttcg ttattaatgt ttgtaattga ctgaatatca acgcttattt
      181 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
      241 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
      301 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
      361 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
      421 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
      481 agtccacgtt cttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
      541 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
      601 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt
      661 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
      721 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
      781 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
      841 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
      901 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
      961 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     1021 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     1081 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     1141 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     1201 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     1261 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     1321 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     1381 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     1441 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     1501 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     1561 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     1621 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     1681 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     1741 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     1801 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     1861 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     1921 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     1981 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     2041 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2101 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     2161 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     2221 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     2281 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     2341 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     2401 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     2461 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     2521 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     2581 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     2641 atagaccagc cgcgtaacct ggcaaaatcg gttacggttg agtaataaat ggatgccctg
     2701 cgtaagcggg tgtgggcgga caataaagtc ttaaactgaa caaaatagat ctaaactatg
     2761 acaataaagt cttaaactag acagaatagt tgtaaactga aatcagtcca gttatgctgt
     2821 gaaaaagcat actggacttt tgttatggct aaagcaaact cttcattttc tgaagtgcaa
     2881 attgcccgtc gtattaaaga ggggcgtggc caagggcatg gtaaagacta tattcgcggc
     2941 gttgtgacaa tttaccgaac aactccgcgg ccgggaagcc gatctcggct tgaacgaatt
     3001 gttaggtggc ggtacttggg tcgatatcaa agtgcatcac ttcttcccgt atgcccaact
     3061 ttgtatagag agccactgcg ggatcgtcac cgtaatctgc ttgcacgtag atcacataag
     3121 caccaagcgc gttggcctca tgcttgagga gattgatgag cgcggtggca atgccctgcc
     3181 tccggtgctc gccggagact gcgagatcat agatatagat ctcactacgc ggctgctcaa
     3241 acttgggcag aacgtaagcc gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa
     3301 gcgcgatgaa tgtcttacta cggagcaagt tcccgaggta atcggagtcc ggctgatgtt
     3361 gggagtaggt ggctacgtct ccgaactcac gaccgaaaag atcaagagca gcccgcatgg
     3421 atttgacttg gtcagggccg agcctacatg tgcgaatgat gcccatactt gagccaccta
     3481 actttgtttt agggcgactg ccctgctgcg taacatcgtt gctgctgcgt aacatcgttg
     3541 ctgctccata acatcaaaca tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga
     3601 ggcatagact gtacaaaaaa acagtcataa caagccatga aaaccgccac tgcgccgtta
     3661 ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg agcgcatacg ctacttgcat
     3721 tacagtttac gaaccgaaca ggcttatgtc aactgggttc gtgccttcat ccgtttccac
     3781 ggtgtgcgtc acccggcaac cttgggcagc agcgaagtcg aggcatttct gtcctggctg
     3841 gcgaacgagc gcaaggtttc ggtctccacg catcgtcagg cattggcggc cttgctgttc
     3901 ttctacggca aggtgctgtg cacggatctg cccttgcttc aggagatcgg tagacctcgg
     3961 ccgtcgcggc gcttgccggt ggtgctgacc ccggatgaag tggttcgcat cctcggtttt
     4021 ctggaaggcg agcatcgttt gttcgcccag gactctagct atagttctag tggttggcta
     4081 cagctttgtt tgtactatca acaggttgaa ctgctgatca acagatcctc tacgcggccg
     4141 cggtaccata acttcgtata gcatacatta tacgaagtta tctggtttaa acgtacccgt
     4201 agtggctatg gcagggcttg ccgccccgac gttggctgcg agccctgggc cttcacccga
     4261 acttgggggt tggggtgggg aaaaggaaga aacgcgggcg tattggtccc aatggggtct
     4321 cggtggggta tcgacagagt gccagccctg ggaccgaacc ccgcgtttat gaacaaacga
     4381 cccaacaccc gtgcgtttta ttctgtcttt ttattgccgt catagcgcgg gttccttccg
     4441 gtattgtctc cttccgtgtt tcagttagcc tcccccatct cccggtaccg catgctatgc
     4501 atcagctgct agcaccatgg ctcgagatcc cgggtgatca agtcttcgtc gagtgattgt
     4561 aaataaaatg taatttacag tatagtattt taattaatat acaaatgatt tgataataat
     4621 tcttatttaa ctataatata ttgtgttggg ttgaattaaa ggtccgtata ctagtatcga
     4681 ttcgcgacct actccggaat attaatagat catggagata attaaaatga taaccatctc
     4741 gcaaataaat aagtatttta ctgttttcgt aacagttttg taataaaaaa acctataaat
     4801 attccggatt attcataccg tcccaccatc gggcgcggat ctacaaccat gtcgtactac
     4861 catcaccatc accatcacga ttacgatatc ccaacgaccg aaaacctgta ttttcagggc
     4921 gccatgggat ccatgagtgc gattaagcca gacatgaaga tcaaactccg tatggaaggc
     4981 aacgtaaacg ggcaccactt tgtgatcgac ggagatggta caggcaagcc ttttgaggga
     5041 aaacagagta tggatcttga agtcaaagag ggcggacctc tgccttttgc ctttgatatc
     5101 ctgaccactg cattccatta cggcaacagg gtattcgcca aatatccaga caacatacaa
     5161 gactatttta agcagtcgtt tcctaagggg tattcgtggg aacgaagctt gactttcgaa
     5221 gacgggggca tttgcaacgc cagaaacgac ataacaatgg aaggggacac tttctataat
     5281 aaagttcgat tttatggtac caactttccc gccaatggtc cagttatgca gaagaagacg
     5341 ctgaaatggg agccctccac tgagaaaatg tatgtgcgtg atggagtgct gacgggtgat
     5401 attgagatgg ctttgttgct tgaaggaaat gcccattacc gatgtgactt cagaactact
     5461 tacaaagcta aggagaaggg tgtcaagtta ccaggcgccc actttgtgga ccactgcatt
     5521 gagattttaa gccatgacaa agattacaac aaggttaagc tgtatgagca tgctgttgct
     5581 cattctggat tgcctgacaa tgccagacga ggtggcggtg gcagcatggc cgaggcaggg
     5641 gctgggctga gcgagaccgt cactgagaca acggttaccg tgacaaccga gcccgagaac
     5701 cggagcctta ccatcaaact tcggaaacgg aagccagaga aaaaggtaga atggacaagt
     5761 gacactgtgg acaatgaaca catgggccgc cgctcatcca aatgctgctg tatttatgag
     5821 aaacctcggg cctttggcga gagctccacg gaaagtgatg aggaggaaga agagggctgt
     5881 ggtcatacac actgtgtacg tggccaccgc aaaggacggc gtcgtgcaac cctaggaccg
     5941 acccccacca cccctcccca gcctcctgac ccttcccagc cccctccagg gccaatgcag
     6001 cactaatgag tcgacgagct cactagtcgc ggccgctttc gaatctagag cctgcagtct
     6061 cgacaagctt gtcgagaagt actagaggat cataatcagc cataccacat ttgtagaggt
     6121 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     6181 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     6241 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     6301 catcaatgta tcttatcatg tctggatctg atcactgctt gagcctagaa gatccggctg
     6361 ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactatcat
     6421 aacccctagg agatccgaac cagataagtg aaatctagtt ccaaactatt ttgtcatttt
     6481 taattttcgt attagcttac gacgctacac ccagttccca tctattttgt cactcttccc
     6541 taaataatcc ttaaaaactc catttccacc cctcccagtt cccaa
//
